Estradiol and estrogen receptor-dependent stabilization of a minivitellogenin mRNA lacking 5,100 nucleotides of coding sequence.
We have developed a transfection assay to investigate the estrogen-mediated stabilization of cytoplasmic vitellogenin mRNA. A minivitellogenin (MV5) gene containing the 5' and 3' untranslated and coding regions but lacking 5,075 nucleotides of internal coding sequence was constructed. Cotransfection of the MV5 plasmid and a Xenopus estrogen receptor expression plasmid into Xenopus liver tissue culture cells yielded a 529-nucleotide MV5 mRNA, which was specifically stabilized by estrogen. MV5 mRNA exhibited the increased stability indicative of positive regulation when the estradiol-estrogen receptor complex was present and was not destabilized by unliganded estrogen receptor. Transfected estrogen receptor, estradiol, and 529 nucleotides of the 5,604-nucleotide vitellogenin B1 mRNA were sufficient for stabilization.
Nielsen
D A
DA
Department of Biochemistry, University of Illinois, Urbana 61801.
Shapiro
D J
DJ
D000818:Animals; D003001:Cloning, Molecular; D004958:Estradiol / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D009865:Oocytes / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics*; D011960:Receptors, Estrogen / Q000502:physiology*; D015183:Restriction Mapping; D013329:Structure-Activity Relationship; D014162:Transfection; D014819:Vitellogenins / Q000235:genetics*; D014982:Xenopus laevis
D012333:RNA, Messenger; D011960:Receptors, Estrogen; D014819:Vitellogenins; D004958:Estradiol